Increase In Glutathione Reductase Activity Research Articles

Dietary exposure to honeybush (Cyclopia) polyphenol-enriched extracts altered redox status and expression of oxidative stress and antioxidant defense-related genes in rat liver

Interest in Cyclopia spp. (honeybush) as a source material for production of polyphenol-enriched extracts (PEEs) for the food ingredient and nutraceutical markets requires investigation of their safety. PEEs of Cyclopia subternata (PECsub) and Cyclopia genistoides (PECgen) were fed (2.5g/kg feed) to male Fischer rats for 28days, while PECsub, having the highest total polyphenol content and antioxidant activity, was also fed for 90days. Their dietary intake did not significantly (P≥0.05) affect body weight gain or relative liver and kidney weight. PECsub resulted in a significant (P<0.05) increase in the total serum bilirubin after 28days, while serum alkaline phosphatase levels were only significantly (P<0.05) increased after 90days, suggesting alterations in the biliary system. No effect on serum iron levels was noticed after 28days, but PECsub significantly (P<0.05) reduced serum iron after 90days. Both PECsub and PECgen increased glutathione reductase activity in the liver significantly (P<0.05) after 28days. Reduced glutathione (GSH) content was significantly (P<0.05) decreased after 90days by PECsub resulting in a marked, but not significant (P≥0.05) decrease in the GSH/GSSG ratio. Considering the expression of oxidative stress and antioxidant defense-related genes after 28days, the expression of eleven genes was altered by PECsub and PECgen. Seven genes, mutually affected by PECsub and PECgen, included (i) antioxidant-related genes, prostaglandin-endoperoxide synthase 1 (Ptgs1), kinesin family member 9 (Kif9) and serine (or cysteine) peptidase inhibitor clade B member 1b (Serpinb1b); (ii) genes involved in reactive oxygen species (ROS) metabolism, xeroderma pigmentosum complementation group A (Xpa) and thioredoxin interacting protein (Txnip) as well as (iii) oxygen transporter-related genes, Fanconi anemia complementation group C (Fancc) and vimentin (Vim). The expression of NADPH oxidase organiser 1 (Noxo1) increased 19.97-fold by PECsub and the thyroid peroxidase (Tpo) 372-fold by PECgen. Changes in the expression of the oxidative stress and antioxidant defense-related genes could be indicative of an underlying stress effected by the honeybush PEEs in the liver. Differential gene expression could likely be attributed to the differences in phenolic composition of the extracts. The daily dietary intake of total polyphenol and individual phenolic constituents, recalculated to a human equivalent dose, was much higher, especially for C. subternata, than normally consumed when habitually drinking “fermented” honeybush herbal tea. Subsequent studies verifying the protein-associated changes governing the oxidative status in the liver is required.

Silicon ameliorates chromium toxicity through phytochelatin-mediated vacuolar sequestration in the roots of Oryza sativa (L.)

ABSTRACTHigh chromium (Cr) in rice causes reduced yield and health hazards. This work investigates how Si alleviates Cr toxicity in rice. Addition of Si under Cr stress restored the growth parameters, total protein content, and membrane stability along with reduced Cr content in shoots, confirming that Si plays critical roles in Cr detoxification in rice. However, Si supplementation under Cr stress caused no significant changes in root Cr content but decreased shoot Cr concentrations compared with Cr-stressed plants, indicating that alleviation of Cr toxicity might be associated with Cr sequestration in roots. Further, concentration of Fe and expression of Fe transporter (OsIRT1) showed no significant changes due to Si supplementation under Cr stress, implying that Fe regulation is not involved with Si-mediated mitigation of Cr toxicity in rice. Further, phytochelatin accumulation and OsPCS1 (phytochelatin synthase) transcripts strongly induced due to the dual treatment of Si and Cr compared with Cr-stressed plants, suggesting that phytochelatin might bind to Cr, which leads to vacuolar sequestration in roots. Furthermore, increased glutathione reductase activity in roots implies that active involvement of ROS scavenging partially ameliorates Cr toxicity in rice plants. The study illustrates first evidences on the effect of Si alleviating Cr toxicity in rice plants.

Effect of temperature on oxidative stress induced by lead in the leaves of Plantago major L.

Abstract Fluctuation of the summer day-time temperatures in the mid-latitudes in a range from 16 to 30°C should not have irreversible negative effects on plants, but may influence metabolic processes including the oxidative stress. To test the effect of moderately high temperature on oxidative stress induced by lead in the leaves of Plantago major L.; the plants were incubated in a water solution of 0, 150, 450, and 900 μM Pb (NO3)2 at 20 and 28°C. Plant reactions were evaluated by the content of thiobarbituric acid reactive substances and ascorbate peroxidase and glutathione reductase activities in leaves after 2, 24, 48, and 72 h. The Pb concentration in the leaves rose with the increase in the Pb content and was higher at 20°C. The increase in stomatal resistance caused by Pb was higher at 28°C. The contents of TBARS increased after 2 h of plant exposure to Pb and the increase was the highest at 900 μM Pb, 28°C. The AsP activity increased up to 50% after 24 h of Pb-treatment at 28°C; the highest increase in glutathione reductase activity was observed after 72 h at 20°C. Thus, the moderately high temperature 28°C compared with optimal 20°C caused a decrease in Pb accumulation in Plantago leaves but amplified the negative effects of lead, especially in the beginning of stress development.

Niacin and Selenium Attenuate Sepsis-Induced Lung Injury by Up-Regulating Nuclear Factor Erythroid 2-Related Factor 2 Signaling.

To determine whether the combination therapy of niacin and selenium attenuates lung injury and improves survival during sepsis in rats and whether its benefits are associated with the activation of the glutathione redox cycle and up-regulation of nuclear factor erythroid 2-related factor 2. Prospective laboratory study. University laboratory. Human lung microvascular endothelial cells and male Sprague-Dawley rats (n = 291). In lipopolysaccharide-exposed cells, the dose-related effects of niacin and selenium were assessed, and the therapeutic effects of the combination therapy of niacin (0.9 mM) and selenium (1.5 μM) were evaluated. The role of nuclear factor erythroid 2-related factor 2 was determined using nuclear factor erythroid 2-related factor 2 knockdown cells. In endotoxemic and cecal ligation and puncture with antibiotics rats, the therapeutic effects of the posttreatments of clinically relevant doses of niacin (360 mg/kg) and selenium (60 μg/kg) were evaluated. Combination therapy reduced the hydrogen peroxide level via the synergistic activation of the glutathione redox cycle, which involves niacin-induced increases in glutathione reductase activity, and reduced the glutathione level and a selenium-induced increase in glutathione peroxidase activity. Combination therapy contributed to the up-regulation of nuclear factor erythroid 2-related factor 2, enhancement of glutathione synthesis, and down-regulation of nuclear factor κB signaling, but nuclear factor erythroid 2-related factor 2 knockdown inhibited the enhancement of glutathione synthesis and down-regulation of the nuclear factor κB pathway. The therapeutic effects of combination therapy on endotoxemic rats were consistent with those on lipopolysaccharide-exposed cells. In addition, the posttreatment of combination therapy attenuated lung injury and improved survival in endotoxemic and cecal ligation and puncture with antibiotics rats. However, individual therapies of niacin or selenium failed to achieve these benefits. The combination therapy of niacin and selenium attenuated lung injury and improved survival during sepsis. Its therapeutic benefits were associated with the synergistic activation of the glutathione redox cycle, reduction of hydrogen peroxide level, and up-regulation of nuclear factor erythroid 2-related factor 2.

Asada-Halliwell pathway maintains redox status in Dioscorea alata tuber which helps in germination

Reactive Oxygen Species (ROS) are important regulatory molecules governing physiological processes. In the present study a biochemical and proteome level comparison of two contrasting growth stages of Dioscorea alata tuber namely germinating and mature tuber was performed in order to understand the tuber physiology and biochemistry. Existence of all the component enzymes [APx (ascorbate peroxidase), GR (glutathione reductase), DHAR (dehydroascorbate reductase), MDHAR (mono-dehydroascorbate reductase)] and major products [ascorbate (ASC) and glutathione (GSH)] of the cycle showed an operational Asada-Halliwell cycle in the tuber. A 2.65 fold increase in ASC content & a 3.8 fold increase in GR activity fortified the redox milieu during germination. In contrast a 5 fold higher H2O2 content (due to 3.08 fold lower APx activity) and accumulation of reactive nitrogen species (RNS) such as nitric oxide (NO, 2.4-fold) and S-nitrosothiol (SNO, 2.08 fold) contributed to overall oxidative conditions in the mature tuber. The carbonic anhydrase (CA, 7.5 fold), DHAR (5.31 fold) and MDHAR (7 fold) activities were higher in the germinating tuber in comparison with the mature tuber. GSNO negatively regulated the CA (3.6 & 3.95 fold), MDHAR (7.5 & 1.5 fold) and APx (2.3 & 1.81 fold) while another NO donor, CysNO negatively regulated the DHAR (2.24 & 1.32 fold) activity in the mature and germinating stages respectively indicating again that the lesser inhibition by NO (via nitrosylation) may be because of overall reducing environment in the germinating tuber. Increased SNO leading to S-nitrosylation of dioscorin was confirmed by Biotin switch assay. This is the first report showing dioscorin nitrosylation. The present analysis showed differential redox regulation and also suggests the physiological relevance of CA, DHAR, MDHAR, APx & GR in tuber germination for the first time. These enzymes may be used as potential markers of tuber germination in future.

Aerobic Training Increased Glutathione Reductase Activity In Adults With Moderate Psoriasis

PURPOSE:. Recent studies have reported that oxidative stress may play a key role in the pathogenesis of psoriasis. In fact, it has been proposed as a potential theraputic target. Accordingly, this study was conducted to determine the influence of aerobic training on antioxidant enzyme activity in adults with psoriasis. METHODS: A total of 100 sedentary, male-adults with moderate psoriasis, according to the PASI (psoriasis area and severity index) score, volunteered for this interventional study. Psoriatic arthritis was considered as exclusión criteria. Participants were randomly allocated to the intervention (n=50) or control group (n=50) using a concealed method. The intervention consisted of a supervised 12-week treadmill training program, 3 sessions/week. According to this protocol, they performed a warm-up, 40-50 min treadmill exercise (increasing 5-minutes each 4-weeks) at a work intensity of 55-65% of peak heart rate (increasing 5% each 4-weeks) and cooling-down. Erythrocyte glutathione reductase (GR, EC 1.6.4.2) activity was measured spectrophotometrically by following the decrease in absorbance due to the oxidation of NADPH. Lastly, the current protocol was approved by an Institutional Ethics Committee. RESULTS: When compared to baseline, aerobic training significantly increased erythrocyte GR activity (8.8±1.0vs10.2±1.3U/gHb;p=0.028) in the intervention group. On the contrary, no significant changes were found in the control group. CONCLUSION: A 12-week aerobic training programme increased antioxidant enzyme activity in adults with psoriasis. While current results are promising, future studies are still required to consolidate this approach in clinical application.

Antioxidant Defense System of Tadpoles (Eupemphix nattereri) Exposed to Changes in Temperature and pH.

Amphibians are highly susceptible to environmental changes, mainly at the larval stage during which they are restricted to small and ephemeral aquatic habitats, which are subject to large fluctuations of abiotic parameters, such as temperature and pH. Consequently, tadpoles experience changes in biochemical, physiological, and molecular processes related to the maintenance of homeostasis, which may lead them to an oxidative stress state. In the present study, we investigated the effects of stress caused by changes in temperature and pH on the antioxidant enzymes catalase (CAT), glucose-6-phosphate dehydrogenase (G6PDH), glutathione reductase (GR) and glutathione-S-transferase (GST) in tadpoles of Eupemphix nattereri. The results show that changes in temperature and pH conditions induce an antioxidant response in tadpoles. GST and GR showed temperature-dependent activities; GST activity was higher in tadpoles exposed to 28°C, whereas GR exhibited increased activity in response to 28°C and 36°C. At 32°C, both GST and GR had the lowest activity. CAT was induced by treatments with acidic (pH 5.0) and alkaline (pH 8.5) pH. Tadpoles exposed to acidic pH also had increased GR activity. The G6PDH was not changed in either experiment. Our data demonstrate that E. nattereri possesses an efficient antioxidant defense system for coping with the damaging effects of heat and acidity/alkalinity conditions in water. The alterations in antioxidant enzymes are probably a result of immediate physiological adaptation of individuals in response to increased production of ROS under environmental stress conditions.

Decreased expression of heme oxygenase is associated with depressive symptoms and may contribute to depressive and hypertensive comorbidity

Background: There is strong evidence that hypertension and depression are comorbid and oxidative stress is implicated in both pathologies. We aimed to elucidate the relationship between biochemical markers of the antioxidant-pro-oxidant equilibrium and depression in hypertension.Methods: Blood was collected from patients diagnosed with depression, hypertension, or comorbid depression and hypertension and healthy age- and sex-matched controls. Whole blood reduced glutathione, erythrocyte superoxide dismutase (SOD-1), glutathione peroxidase (GPx-1), glutathione reductase (GR), malondialdehyde (MDA), and plasma hydrogen peroxide (H2O2) were assayed using spectrophotometry, and heme oxygenase (HO-1) levels were determined immunoenzymatically.Results: Both hypertension and depression were associated with altered antioxidant-pro-oxidant profiles. Decreased GPx-1 and SOD-1 activities, increased GR activity, increased levels of GSH, and increased concentrations of MDA and H2O2 were observed in patients compared to controls. Inducible HO-1 was specifically decreased in patients with depression and was significantly associated with both the prevalence and severity of depressive symptoms.Conclusions: Heme oxygenase is a biological factor that might explain the relationship between inflammation, oxidative stress, and the biological and functional changes in brain activity in depression. HO-1 is a candidate depression biomarker and provides an avenue for novel preventative and diagnostic strategies against this disease.

Inhibitory Effect of Yongdamsagan-Tang Water Extract, a Traditional Herbal Formula, on Testosterone-Induced Benign Prostatic Hyperplasia in Rats.

Yongdamsagan-tang, a traditional herbal formula, is used widely for the treatment of inflammation and viral diseases. In this study, we investigated whether Yongdamsagan-tang water extract (YSTE) affects testosterone propionate- (TP-) induced benign prostatic hyperplasia (BPH) in a rat model. To induce BPH, rats were injected subcutaneously with 10 mg/kg of TP every day. YSTE was administrated daily by oral gavage at doses of 200 and 500 mg/kg along with the TP injection. After 4 weeks, prostates were collected, weighed, and analyzed. The relative prostrate weight was significantly lower in both YSTE groups (200 and 500 mg/kg/day) compared with the TP-induced BPH group. YSTE administration reduced the expression of proliferation markers PCNA, cyclin D1, and Ki-67 and the histological abnormalities observed in the prostate in TP-induced BPH rats. YSTE attenuated the increase in the TP-induced androgen concentration in the prostate. The YSTE groups also showed decreased lipid peroxidation and increased glutathione reductase activity in the prostate. These findings suggest that YSTE effectively prevented the development of TP-induced BPH in rats through antiproliferative and antioxidative activities and might be useful in the clinical treatment of BPH.

The role of oxidative protein modification and the gluthatione system in modulation of the redox status of breast epithelial cells

The effects of the SH-group blocker N-ethylmaleimide (NEM) and thiol group protector 1,4-dithioerythritol (DTE) on the redox status of cells HBL-100 cells, oxidative modification of their proteins and the state of glutathione and thioredoxin systems have been investigated. Breast epithelial cells cultivated in the presence of NEM were characterized by decreased redox status, increased glutathione reductase activity, and increased concentrations of products of irreversible oxidative modification of protein and amino acids. Cultivation of HBL-100 cells in the presence of DTE resulted in a shift of the redox status towards reduction processes and increased reversible protein modification by glutathionylation. The proposed model of intracellular redox modulation may be used in the development of new therapeutic approaches to treat diseases accompanied by impaired redox homeostasis (e.g. oncologic, inflammatory, cardiovascular and neurodegenerative disease).

The role of heat shock proteins 27 and 70 in redox-dependent regulation of apoptosis in Jurkat tumor cells

Heat shock proteins Hsp) act as molecular chaperones, protecting enzymes and other proteins against reactive oxygen species. The objective of the study was to investigate the role of Hsp27 in maintaining the balance of the glutathione system and Hsp70 concentrations as well as in implementing Jurkat tumor cell apoptosis. Addition of the Hsp27 inhibitor KRIBB3 (5-(5-ethyl-2-hydroxy-4-methoxyphenyl)-4-(4-methoxyphenyl)-isoxazol) to Jurkat cells resulted in glutathione redox imbalance (increased GSSG and increased glutathione reductase activity), a decrease in Hsp70 concentrations, and also increased cell apoptosis as compared with to the intact cell culture. The proposed selective regulation of chaperone activity is a promising direction in regulating apoptosis at the cellular level.

Daily intake of lead in Wistar rats at different ages: Biochemical, genotoxic and physiological effects.

The effects of the daily intake of feed containing lead (2.0 mg Pb kg(-1)) were evaluated in 45 (Pb45) and 90 (Pb90)-day-old Wistar female rats. Compared to the respective control groups, Pb45 rats consumed more feed and showed greater weight gain, but these parameters returned to control values in Pb90 rats. Higher blood glucose levels were observed in both Pb groups, whereas plasma insulin concentrations were higher in Pb45 but lower in the Pb90 group. Liver glycogen content was lower only in the Pb90 rats. There were no changes in plasma cortisol and acetylcholinesterase activity in the brain. Hematological alterations were observed only in Pb90 rats, which showed lower hemoglobin levels. In the liver, Pb45 rats showed decreased catalase and glutathione peroxidase activities and increased glutathione reductase activity, but in the Pb90 group, glutathione levels were higher. Increased hepatic lipid peroxidation and DNA damage in the lymphocytes were observed in both Pb groups. These results indicate that the daily intake of Pb for different periods results in metabolic changes and in the establishment of oxidative and genotoxic damage in female rats.

Diets supplemented with seaweed affect metabolic rate, innate immune, and antioxidant responses, but not individual growth rate in European seabass (Dicentrarchus labrax)

This study investigated the effects of seaweed dietary supplementation on measures of fish performance including aerobic metabolism, digestive enzymes activity, innate immune status, oxidative damage, and growth rate using European seabass (Dicentrarchus labrax). Fish were fed for 49 days with three different diets: a control diet (CTRL), a Gracilaria-supplemented diet (GR7.5), and a mixed diet (Mix) composed of Gracilaria, Fucus, and Ulva genera representatives. All diets were isoenergetic (22 kJ g−1 adjusted for dry matter (DM)), isoproteic (47 %DM), and isolipidic (18 %DM) and tested in triplicate groups of 20 fish (initial body weight 25.5 ± 4.1 g). Final results showed similar growth rates and digestive activities between diets. Maximum and standard metabolic rates and aerobic metabolic scope revealed comparable results for the three diets. In contrast, fish fed with GR7.5 exhibited elevated routine metabolic rate (190.7 mg O2 kg−1 h−1). Fish fed with the GR7.5 and Mix diets had lower alternative complement pathway (ACH50) (62.5 and 63 units mL−1 respectively) than CTRL (84 units mL−1) GR7.5 increased lipid peroxidation and cholinesterase levels, as well as glutathione s-transferase activity. Mix diet increased glutathione reductase activity when compared to CTRL. Collectively, our findings suggest that dietary seaweed supplementation may alter seabass metabolic rate, innate immune, and antioxidant responses without compromising growth parameters.

The Alleviation of the Adverse Effects of Salt Stress in the Tomato Plant by Salicylic Acid Shows A Time- and Organ-Specific Antioxidant Response

Abstract In the last decade contradictory results have been published as to whether exogenous salicylic acid (SA) can increase salt stress tolerance in cultivated plants by inducing an antioxidant response. Salt stress injury in tomato was mitigated only in cases when the plant was hardened with a high concentration of SA (~10−4 M), low concentrations were ineffective. An efficient accumulation of Na+ in older leaves is a well-known response to salt stress in tomato plants (Solanum lycopersicum cv. Rio fuego) but it remains largely unexplored whether young and old leaves or root tissues have a distinct antioxidant status during salt stress after hardening with 10−7 M or 10−4 M SA. The determination of superoxide dismutase (SOD) and catalase (CAT) activity revealed that the SA-induced transient increases in these enzyme activities in young leaf and/or root tissues did not correlate with the salt tolerance of plants. Salt stress resulted in a tenfold increase in ascorbate peroxidase (APX) activities of young leaves and significant increases in APX and glutathione reductase (GR) activities of the roots hardened with 10−4 M SA. Both total ascorbate (AsA) and glutathione pools reached their highest levels in leaves after 10−7 M SA pre-treatment. However, in contrast to the leaves, the total pool of AsA decreased in the roots under salt stress and thus, due to low APX activity, active oxygen species were scavenged by ascorbate non-enzymatically in these tissues. The increased GR activities in the roots after treatment with 10−4 M SA enabled plants to enhance the reduced glutathione (GSH) pool and maintain the redox status of AsA under high salinity, which led to increased salt tolerance.

Redox proteomic analysis of mytilus edulis gills: effects of the pharmaceutical diclofenac on a non-target organism.

Veterinary and human pharmaceuticals are an emerging category of chemical pollutants with potential to cause serious toxicity to non-target organisms. Filter-feeding aquatic organisms such as mussels are especially threatened. In this study, the blue mussel, Mytilus edulis, was exposed to two doses (0.2 mg/L and 1 mg/L) of the anti-inflammatory diclofenac. Effects on the gill, the principal feeding organ of mussels, were investigated. It was noted that, while no effect was evident on gill glutathione transferase or catalase activities, there was a tissue-specific increase in glutathione reductase activity and reduction in total protein thiol groups. Two dimensional electrophoresis was performed and some affected proteins identified by in-gel tryptic digestion and peptide mass fingerprinting. Of these, four unique proteins (caspase 3/7-4, heat-shock cognate protein 70, a predicted enolase-like protein, arginine kinase) were found to be oxidized whilst eight unique proteins (β-tubulin, actin, isocitrate dehydrogenase, arginine kinase, heavy metal-binding HIP, cytosolic malate dehydrogenase, proteasome subunit alpha type 2, Mg: bb02e05 (glyceraldehyde-3-phosphate dehydrogenase) and superoxide dismutase) were found to have altered abundance. In addition, bioinformatic analysis suggested putative identities for six hypothetical proteins which either were oxidized or decreased in abundance. These were; 78 kDa glucose-regulated protein precursor, α-enolase, calreticulin, mitochondrial H + -ATPase, palmitoyl protein thioesterase 1 and initiation factor 5a. It is concluded that diclofenac causes significant oxidative stress to gills and that this affects key structural, metabolic and stress-response proteins.

Glutathione metabolism system under condition of transplantation of muscle tissue in rats

The paper presents the results of determining the amount of reduced glutathione, glutathione reductase and glutathione peroxidase activity at transplantation of muscle tissue taken from rats from the same litter. During the work 2 surgical interventions were carried out: transplantation of muscle tissue taken from the same litter of rats, and operation without replanting. Transplantation was performed on nonlinear white male rats weighing 180–300 g. The animals were taken out of the experiment by passing electric current through the medulla oblongata. For donor transplants males from the same litter were used. In adult donor rats abdominal muscle tissue was extracted and transplanted to homologous tissue of the recipient rat. The same procedure was carried out with femoral muscle tissue. Operation without replanting was used to compare the effects of changes in the number of investigated parameters of the surgical exposure. Tissue not subject to any surgery served as a control. Parameters under study were determined on the first, third and seventh day after transplantation. Glutathione levels in tissues were determined byE. Butler, A. Dyubona, B. Kelly. Determination of glutathione reductase is based on the reduction of NADPH registration. Glutathione peroxidase activity was determined by accumulation of oxidized glutathione. Surgery is known to lead to expression of oxidative stress in the organs operated. Special role in antioxidant protection of the body is given to thiols. Expressed hydrophilic properties provide their high content in water fraction of cells and ability to protect biologically important molecules (enzymes, nucleic acids, hemoglobin) from oxidative damage. Property of thiol compounds is known to inhibit both fermented and not fermented free radical oxidation. For the first time it was found that transplantation of muscle tissue taken from animals from the same litter lead to significant reduction of glutathione in the femoral muscle of the recipient and abdominal muscle tissue of the donor and recipient and to significant increase in glutathione reductase activity in the femoral muscle tissue of the donor and in the abdominal muscle tissue of the donor and recipient.

Redox proteomics changes in the fungal pathogen Trichosporon asahii on arsenic exposure: identification of protein responses to metal-induced oxidative stress in an environmentally-sampled isolate.

Trichosporon asahii is a yeast pathogen implicated in opportunistic infections. Cultures of an isolate collected from industrial wastewater were exposed for 2 days to 100 mg/L sodium arsenite (NaAsO2) and cadmium (CdCl2). Both metals reduced glutathione transferase (GST) activity but had no effect on superoxide dismutase or catalase. NaAsO2 exposure increased glutathione reductase activity while CdCl2 had no effect. Protein thiols were labeled with 5-iodoacetamido fluorescein followed by one dimensional electrophoresis which revealed extensive protein thiol oxidation in response to CdCl2 treatment but thiol reduction in response to NaAsO2. Two dimensional electrophoresis analyses showed that the intensity of some protein spots was enhanced on treatment as judged by SameSpots image analysis software. In addition, some spots showed decreased IAF fluorescence suggesting thiol oxidation. Selected spots were excised and tryptic digested for identification by MALDI-TOF/TOF MS. Twenty unique T. asahii proteins were identified of which the following proteins were up-regulated in response to NaAsO2: 3-isopropylmalate dehydrogenase, phospholipase B, alanine-glyoxylate aminotransferase, ATP synthase alpha chain, 20S proteasome beta-type subunit Pre3p and the hypothetical proteins A1Q1_08001, A1Q2_03020, A1Q1_06950, A1Q1_06913. In addition, the following showed decreased thiol-associated fluorescence consistent with thiol oxidation; aconitase; aldehyde reductase I; phosphoglycerate kinase; translation elongation factor 2; heat shock protein 70 and hypothetical protein A1Q2_04745. Some proteins showed both increase in abundance coupled with decrease in IAF fluorescence; 3-hydroxyisobutyryl- CoA hydrolase; homoserine dehydrogenase Hom6 and hypothetical proteins A1Q2_03020 and A1Q1_00754. Targets implicated in redox response included 10 unique metabolic enzymes, heat shock proteins, a component of the 20S proteasome and translation elongation factor 2. These data suggest extensive proteomic alterations in response to metal-induced oxidative stress in T. asahii. Amino acid metabolism, protein folding and degradation are principally affected.

Effects of passion fruit (Passiflora edulis) byproduct intake in antioxidant status of Wistar rats tissues

Passion fruit (Passiflora edulis) is a native species from Brazil explored in fruit culture. Peels are byproducts of juice industrial processing and are usually discarded. They are source of fibers and other bioactive substances, which may have antioxidant role and to protect cells against oxidative stress. This study aimed to evaluate the antioxidant status of P. edulis peel flour in vivo. The male Wistar rats (77 days old) were divided in Peel and Control groups (n = 4). Control group was fed with standard diet (AIN-93M) and Peel group, received a diet in which fifty percent of the cellulose content was replaced by fiber from PPF. After 15 days, the animals were anaesthetized and sacrificed. Peel group consumed greater amount of diet without alterations in body weight of animals. The experimental group showed no differences in serum antioxidant status (FRAP and TBARS assays) (P > 0.05), but they presented lower antioxidant potential according to ORAC assay (P < 0.05). In the kidneys of Peel group, a reduction in lipid peroxidation by TBARS assay (P < 0.05) was observed. In addition, the liver of the animals fed with PPF showed high TBARS levels, increased glutathione reductase activity (GR), decreased glutathione peroxidased activity (GPx) and superoxide dismutase activity (SOD).

Impact of ovariectomy, high fat diet, and lifestyle modifications on oxidative/antioxidative status in the rat liver.

AimTo estimate the impact of high fat diet and estrogen deficiency on the oxidative and antioxidative status in the liver of the ovariectomized rats, as well as the ameliorating effect of physical activity or consumption of functional food containing bioactive compounds with antioxidative properties on oxidative damage in the rat liver.MethodsThe study was conducted from November 2012 to April 2013. Liver oxidative damage was determined by lipid peroxidation levels expressed in terms of thiobarbituric acid reactive substances (TBARS), while liver antioxidative status was determined by catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), glutathione reductase (GR) activities, and glutathione (GSH) content. Sixty-four female Wistar rats were divided into eight groups: sham operated and ovariectomized rats that received either standard diet, high fat diet, or high fat diet supplemented with cereal selenized onion biscuits or high fat diet together with introduction of physical exercise of animals.ResultsHigh fat diet significantly increased TBARS content in the liver compared to standard diet (P = 0.032, P = 0.030). Furthermore, high fat diet decreased the activities of CAT, GR, and GST, as well as the content of GSH (P < 0.050). GPx activity remained unchanged in all groups. Physical activity and consumption of cereal selenized onion biscuits showed protective effect through increased GR activity in sham operated rats (P = 0.026, P = 0.009), while in ovariectomized group CAT activity was increased (P = 0.018) in rats that received cereal selenized onion biscuits.ConclusionFeeding rats with high fat diet was accompanied by decreased antioxidative enzyme activities and increased lipid peroxidation. Bioactive compounds of cereal selenized onion biscuits showed potential to attenuate the adverse impact of high fat diet on antioxidative status.

Ceramide Transfer Protein Deficiency Compromises Organelle Function and Leads to Senescence in Primary Cells

Ceramide transfer protein (CERT) transfers ceramide from the endoplasmic reticulum (ER) to the Golgi complex. Its deficiency in mouse leads to embryonic death at E11.5. CERT deficient embryos die from cardiac failure due to defective organogenesis, but not due to ceramide induced apoptotic or necrotic cell death. In the current study we examined the effect of CERT deficiency in a primary cell line, namely, mouse embryonic fibroblasts (MEFs). We show that in MEFs, unlike in mutant embryos, lack of CERT does not lead to increased ceramide but causes an accumulation of hexosylceramides. Nevertheless, the defects due to defective sphingolipid metabolism that ensue, when ceramide fails to be trafficked from ER to the Golgi complex, compromise the viability of the cell. Therefore, MEFs display an incipient ER stress. While we observe that ceramide trafficking from ER to the Golgi complex is compromised, the forward transport of VSVG-GFP protein is unhindered from ER to Golgi complex to the plasma membrane. However, retrograde trafficking of the plasma membrane-associated cholera toxin B to the Golgi complex is reduced. The dysregulated sphingolipid metabolism also leads to increased mitochondrial hexosylceramide. The mitochondrial functions are also compromised in mutant MEFs since they have reduced ATP levels, have increased reactive oxygen species, and show increased glutathione reductase activity. Live-cell imaging shows that the mutant mitochondria exhibit reduced fission and fusion events. The mitochondrial dysfunction leads to an increased mitophagy in the CERT mutant MEFs. The compromised organelle function compromise cell viability and results in premature senescence of these MEFs.